Catharanthus hybrid &#39;pas1305707&#39; and parent thereof

ABSTRACT

The invention provides seeds and plants of  Catharanthus  hybrid ‘PAS1305707’ and the parent lines thereof. The invention thus relates to the plants, seeds, plant parts, and tissue cultures of  Catharanthus  hybrid ‘PAS1305707’ and the parent lines thereof, and to methods for producing a  Catharanthus  plant produced by crossing such plants with themselves or with another  Catharanthus  plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates plants, seeds, plant parts and tissue cultures of  Catharanthus  hybrid ‘PAS1305707’ and the parent lines thereof comprising introduced beneficial or desirable traits.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of Catharanthus hybrid ‘PAS1305707’ andthe parent Catharanthus line R2358D.

BACKGROUND OF THE INVENTION

Of the eight known Catharanthus species, seven originate fromMadagascar, and the exception Catharanthus pusillus is endemic to Indiaand Sri Lanka. Of importance is Catharanthus roseus, commonly known asMadagascar periwinkle or vinca, which is valued for production ofmedicinal indole alkaloids. In addition, it is frequently grown annuallyfrom seed or less commonly cuttings in temperate climates for use insummer bedding or as a pot plant. It has long been grown as anornamental in tropical regions of the world. As a consequence of itsability to self-pollinate, Catharanthus roseus is now widely naturalizedin many tropical regions. Hybridization is less common; however, thereare natural hybrids between Catharanthus longifolius and Catharanthusroseus. Man-made crosses have yielded hybrids between Catharanthusovalis and Catharanthus longifolius, and between Catharanthus roseus andCatharanthus trichophyllus.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a Catharanthus plant ofthe hybrid designated ‘PAS1305707’ or Catharanthus line R2358D. Alsoprovided are Catharanthus plants having all the physiological andmorphological characteristics of such a plant. Parts of theseCatharanthus plants are also provided, for example, including a flower,pollen, a leaf, an ovule, an embryo, a seed, and a cell of the plant.

In another aspect of the invention, a plant of Catharanthus hybrid‘PAS1305707’ or Catharanthus line R2358D comprising an added heritabletrait is provided. The heritable trait may comprise a genetic locus thatis, for example, a dominant or recessive allele. In one embodiment ofthe invention, a plant of Catharanthus hybrid ‘PAS1305707’ orCatharanthus line R2358D is defined as comprising a single locusconversion. In specific embodiments of the invention, an added geneticlocus confers one or more traits such as, for example, herbicidetolerance, insect resistance, disease resistance, and modifiedcarbohydrate metabolism. In further embodiments, the trait may beconferred by a naturally occurring gene introduced into the genome of aline by backcrossing, a natural or induced mutation, or a transgeneintroduced through genetic transformation techniques into the plant or aprogenitor of any previous generation thereof. When introduced throughtransformation, a genetic locus may comprise one or more genesintegrated at a single chromosomal location.

In some embodiments, a single locus conversion includes one or moresite-specific changes to the plant genome, such as, without limitation,one or more nucleotide modifications, deletions, or insertions. A singlelocus may comprise one or more genes or nucleotides integrated ormutated at a single chromosomal location. In one embodiment, a singlelocus conversion may be introduced by a genetic engineering technique,methods of which include, for example, genome editing with engineerednucleases (GEEN). Engineered nucleases include, but are not limited to,Cas endonucleases; zinc finger nucleases (ZFNs); transcriptionactivator-like effector nucleases (TALENs); engineered meganucleases,also known as homing endonucleases; and other endonucleases for DNA orRNA-guided genome editing that are well-known to the skilled artisan.

The invention also concerns the seed of Catharanthus hybrid ‘PAS1305707’or Catharanthus line R2358D. The Catharanthus seed of the invention maybe provided, in particular embodiments, as an essentially homogeneouspopulation of Catharanthus seed of Catharanthus hybrid ‘PAS1305707’ orCatharanthus line R2358D. Essentially homogeneous populations of seedare generally free from substantial numbers of other seed. Therefore,seed of hybrid ‘PAS1305707’ or Catharanthus line R2358D may be definedas forming at least about 90% of the total seed, including at leastabout 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more of the seed.The seed population may be separately grown to provide an essentiallyhomogeneous population of Catharanthus plants designated ‘PAS1305707’ orR2358D.

In yet another aspect of the invention, a tissue culture of regenerablecells of a Catharanthus plant of hybrid ‘PAS1305707’ or Catharanthusline R2358D is provided. The tissue culture will preferably be capableof regenerating Catharanthus plants capable of expressing all of thephysiological and morphological characteristics of the starting plant,and of regenerating plants having substantially the same genotype as thestarting plant. Examples of some of the physiological and morphologicalcharacteristics of the Catharanthus hybrid ‘PAS1305707’ and Catharanthusline R2358D include those traits set forth in the tables herein. Theregenerable cells in such tissue cultures may be derived, for example,from embryos, meristems, cotyledons, pollen, leaves, anthers, roots,root tips, pistils, flowers, cuttings, seeds, and stems. Still further,the present invention provides Catharanthus plants regenerated from atissue culture of the invention, the plants having all the physiologicaland morphological characteristics of Catharanthus hybrid ‘PAS1305707’ orCatharanthus line R2358D.

In still yet another aspect of the invention, processes are provided forproducing Catharanthus seeds, plants and parts thereof, which processesgenerally comprise crossing a first parent Catharanthus plant with asecond parent Catharanthus plant, wherein at least one of the first orsecond parent Catharanthus plants is a plant of Catharanthus lineR2358D. These processes may be further exemplified as processes forpreparing hybrid Catharanthus seed or plants, wherein a firstCatharanthus plant is crossed with a second Catharanthus plant of adifferent, distinct genotype to provide a hybrid that has, as one of itsparents, a plant of Catharanthus line R2358D. In these processes,crossing will result in the production of seed. The seed productionoccurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent Catharanthusplant, often in proximity so that pollination will occur for example,mediated by insect vectors. Alternatively, pollen can be transferredmanually. Where the plant is self-pollinated, pollination may occurwithout the need for direct human intervention other than plantcultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent Catharanthus plants into plants that bear flowers. A thirdstep may comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent Catharanthus plants. Yet another stepcomprises harvesting the seeds from at least one of the parentCatharanthus plants. The harvested seed can be grown to produce aCatharanthus plant or hybrid Catharanthus plant.

The present invention also provides the Catharanthus seeds and plantsproduced by a process that comprises crossing a first parentCatharanthus plant with a second parent Catharanthus plant, wherein atleast one of the first or second parent Catharanthus plants is a plantof Catharanthus hybrid ‘PAS1305707’ or Catharanthus line R2358D. In oneembodiment of the invention, Catharanthus seed and plants produced bythe process are first generation (F1) hybrid Catharanthus seed andplants produced by crossing a plant in accordance with the inventionwith another, distinct plant. The present invention further contemplatesplant parts of such an F1 hybrid Catharanthus plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F1 hybrid Catharanthus plant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from Catharanthus hybrid ‘PAS1305707’ orCatharanthus line R2358D, the method comprising the steps of: (a)preparing a progeny plant derived from Catharanthus hybrid ‘PAS1305707’or Catharanthus line R2358D, wherein said preparing comprises crossing aplant of the hybrid ‘PAS1305707’ or Catharanthus line R2358D with asecond plant; and (b) crossing the progeny plant with itself or a secondplant to produce a seed of a progeny plant of a subsequent generation.In further embodiments, the method may additionally comprise: (c)growing a progeny plant of a subsequent generation from said seed of aprogeny plant of a subsequent generation and crossing the progeny plantof a subsequent generation with itself or a second plant; and repeatingthe steps for an additional 3-10 generations to produce a plant derivedfrom hybrid ‘PAS1305707’ or Catharanthus line R2358D. The plant derivedfrom hybrid ‘PAS1305707’ or line R2358D may be an inbred line, and theaforementioned repeated crossing steps may be defined as comprisingsufficient inbreeding to produce the inbred line. In the method, it maybe desirable to select particular plants resulting from step (c) forcontinued crossing according to steps (b) and (c). By selecting plantshaving one or more desirable traits, a plant derived from Catharanthushybrid ‘PAS1305707’ or Catharanthus line R2358D is obtained whichpossesses some of the desirable traits of the line/hybrid as well aspotentially other selected traits.

In still yet another aspect of the invention, the genetic complement ofCatharanthus hybrid ‘PAS1305707’ or Catharanthus line R2358D isprovided. The phrase “genetic complement” is used to refer to theaggregate of nucleotide sequences, the expression of which sequencesdefines the phenotype of, in the present case, a Catharanthus plant, ora cell or tissue of that plant. A genetic complement thus represents thegenetic makeup of a cell, tissue or plant, and a hybrid geneticcomplement represents the genetic makeup of a hybrid cell, tissue orplant. The invention thus provides Catharanthus plant cells that have agenetic complement in accordance with the Catharanthus plant cellsdisclosed herein, and plants, seeds and plants containing such cells.Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that Catharanthus hybrid ‘PAS1305707’ or Catharanthusline R2358D could be identified by any of the many well-known techniquessuch as, for example, Simple Sequence Length Polymorphisms (SSLPs)(Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), RandomlyAmplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs), ArbitraryPrimed Polymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by Catharanthus plant cells,tissues, plants, and seeds, formed by the combination of a haploidgenetic complement of a Catharanthus plant of the invention with ahaploid genetic complement of a second Catharanthus plant, preferably,another, distinct Catharanthus plant. In another aspect, the presentinvention provides a Catharanthus plant regenerated from a tissueculture that comprises a hybrid genetic complement of this invention.

In still yet another aspect, the invention provides a method ofdetermining the genotype of a plant of Catharanthus hybrid ‘PAS1305707’or Catharanthus line R2358D comprising detecting in the genome of theplant at least a first polymorphism. The method may, in certainembodiments, comprise detecting a plurality of polymorphisms in thegenome of the plant. The method may further comprise storing the resultsof the step of detecting the plurality of polymorphisms on a computerreadable medium. The invention further provides a computer readablemedium produced by such a method.

In one aspect, the invention provides a method of producing aCatharanthus seed, the method comprising (a) obtaining a plant of thepresent invention, wherein the plant has been cultivated to maturity;and (b) collecting a Catharanthus seed from the plant.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds, and derivatives of Catharanthus hybrid ‘PAS1305707’ andCatharanthus line R2358D.

The hybrid ‘PAS1305707’ was produced by the cross of parent lines R2358Dand M6948D. The parent lines show uniformity and stability within thelimits of environmental influence. By crossing the parent lines, uniformplants of hybrid ‘PAS1305707’ can be obtained.

A. Origin and Breeding History of Catharanthus Hybrid ‘PAS1305707’

The parents of hybrid ‘PAS1305707’ are R2358D and M6948D with R2358Dused as the female parent. These parents were created as follows:

Catharanthus hybrid ‘PAS1305707’ originated from a cross in 2014 betweentwo proprietary inbred lines. The cross was made between parental linesR2358D×M6948D in a greenhouse in Guadalupe, Calif. In 2014 the hybridwas trialed in California and Florida. In both locations the hybrid wasevaluated for general horticultural characters. At a research stationlocated near Ruskin, Fla. the hybrid was evaluated for generalhorticultural characters and field resistance to disease.

The hybrid cross was repeated in 2015, and seed was sent to researchstations in California, Illinois, and Florida for repeat evaluation. Thehabit, color, and other horticultural characters were uniform andstable.

In 2015, a large production test was conducted in Guatemala to evaluateseed yield and seed quality of the hybrid. Seed from this productiontest was sent to research stations in California, Illinois, and Texasfor evaluation in 2016. The hybrid was designated ‘PAS1305707’ and hasshown uniformity and stability, as described in the cultivar descriptioninformation.

B. Origin and Breeding History of Catharanthus Female Inbred Line R2358D

The inbred line R2358D was developed using the pedigree breeding system.The original developmental cross was made in the greenhouses inGuadalupe, Calif. in 2009 using two proprietary inbred lines09-0592×09-0949.

The resulting F₁ from this cross was sown in a greenhouse in 2010 andbulk-sibbed to produce F₂ seed. In 2011, the F₂ population wasevaluated, and single plant selections were made for generalhorticultural characters, including a well branched habit, largeflowers, and disease resistance.

Several F₃ families were selected in 2012 that were the most uniform forthe above characters. Single plant selections were made to produce F₄families. Under Florida growing conditions, F₃ families were alsoevaluated for horticultural characters and field resistance to disease.

Subsequent F₄ to F₅ generations were advanced from 2012 through 2014using self-pollinated single plant selections, while continuing toselect families with the most uniform previously described characters.One of the F₅ families was massed in 2014 to produce breeder's seed. In2015, the massed seed from 2014 was used to make stock seed.

The plants massed in 2014 and 2015 were uniform and stable. No variantsor off-types were observed in either the breeder's seed increase or thestock seed increase.

The inbred line was designated R2358D. It was stable and uniform aftertwo generations of seed increase, which included the breeder's seedincrease and the stock seed increase.

C. Origin and Breeding History of Catharanthus Male Inbred Line M6948D

The inbred line M6948D was developed using the pedigree breeding system.The original germplasm dates back to 2003 from a proprietary inbreddeveloped from earlier vinca breeding. The F₆ inbred was designated03-070 and was developed in the greenhouses in Guadalupe, Calif.

In 2008, the F₆ inbred line 03-070 was sown in the Guadalupe greenhousesand was evaluated under Florida growing condition for horticulturalcharacters and field resistance to disease. Single plant selections weremade in the greenhouse and advanced to the F₇ generation.

In 2009, single plant selections were again evaluated and single plantsselections were made and self-pollinated to produce F₈ families.

One of the F₈ families was massed in 2010 to produce breeder's seed. In2011, the massed seed from 2010 was used to make stock seed.

The plants massed in 2010 and the plants massed in 2011 were uniform andstable. No variants or off-types were observed in either the breeder'sseed increase or the stock seed increase.

The inbred line was designated M6948D. It was stable and uniform aftertwo generations of seed increase, which included the breeder's seedincrease and the stock seed increase.

D. Physiological and Morphological Characteristics of CatharanthusHybrid ‘PAS1305707’ and Catharanthus Line R2358D

The new hybrid cultivar ‘PAS1305707’ and female inbred line R2358D havenot been observed under all possible environmental conditions to date.As is known in the art, it is possible that the phenotypes may varysomewhat with variations in the environment, such as temperature, lightintensity, and day length, without, however, any variance in genotype.

The following descriptions and measurements describe plants producedfrom seed and grown in a glass-covered greenhouse under conditionscomparable to those used in commercial practice. The plants were grownutilizing a soilless growth medium in one-gallon containers for 21 weeksin Guadalupe, Calif. Greenhouse temperatures were maintained atapproximately 75° F. to 85° F. (24° C. to 30° C.) during the day andapproximately 65° F. to 68° F. (18° C. to 20° C.) during the night. Nosupplemental lighting was provided.

The chart used in the identification of colors described herein is thePantone Plus Series Color Bridge, 2nd Edition, except where generalcolor terms of ordinary significance are used. The color values weredetermined in February 2020 under natural light conditions in Guadalupe,Calif. Measurements and numerical values represent averages of typicalplants.

A description of the physiological and morphological characteristics ofthe plants described herein are presented in the tables that follow.

TABLE 1 Physiological and Morphological Characteristics of Hybrid‘PAS1305707’ CHARACTERISTIC ‘PAS1305707’ ‘PAS1053045’ 1. OVERALL PLANTHABIT (at flowering stage): Species: 1 = C. roseus; 2 = Other 1 1Ploidy: 1 = Haploid; 2 = Diploid; 3 = Triploid; 4 = Tetrapioid 2 2 LifeCycle: 1 = Annual; 2 = Biennial; 3 = Perennial 1 1 Growth Habit: 1 =Determinate; 2 = Semi-determinate; 2 2 3 = Indeterminate Growth Form: 1= Upright; 2 = Semi-prostrate; 3 = Prostrate 1 1 Flowering: 1 = VeryEarly; 2 = Early; 3 = Mid-season; 4 = Late; 2 2 5 = Continuous Days fromPlanting to 50% Flowering 56 68 Length of Flowering Season in Days Untilfrost Until frost Plant Height at Maturity (cm) 23.7 28.6 Plant Width atMaturity (cm) 29.4 40.1 Plant Height Class: 1 = Extra Dwarf; 2 = Dwarf;3 = Semi-dwarf; 3 3 4 = Tall Plant Width Class: 1 = Compact; 2 =Semi-compact; 2 2 3 = Spreading/Lax 2. STEM: Profile: 1 = Straight; 2 =Zig-Zag 1 1 Branching Pattern: 1 = Single Stem; 2 = Few Branches; 3 =Many 3 3 Branches Stem Length from Base of Stem to Terminal Flower (cm)24.6 28.1 Number of Internodes Below First Branch 1 1 Number of FirstOrder Branches (From Main Stem) 14 13 Stem Anthocyanin: 1 = Absent; 2 =Along Veins Only; 3 = Solid 1 1 Coloration 3. FOLIAGE: Leaf Type: 1 =Simple; 2 = Compound 1 1 Leaf Margin: 1 = Entire; 2 = Serrate; 3 = Other1 1 Leaf Odor: 1 = None; 2 = Mild; 3 = Strong 1 1 Petiole Anthocyanin: 1= Absent; 2 = Mild; 3 = Strong 1 1 Leaf Shape: 1 = Lanceolate; 2 =Elliptic; 3 = Obovate; 4 = Ovate 2 2 Leaf Width (mm) 34.2 29.1 LeafLength (mm) 87.1 106.2 LEAF DORSAL SIDE: Leaf Color: 1 = Light Green; 2= Medium Green; 3 = Dark Green; 3 3 4 = Other Color Chart Code  574C 574C Pubescence: 1 = Absent; 2 = Light; 3 = Heavy 1 1 Luster: 1 = Dull;2 = Shiny 1 1 LEAF VENTRAL SIDE: Leaf Color: 1 = Light Green; 2 = MediumGreen; 3 = Dark Green; 2 2 4 = Other Color Chart Code  371C  371CPubescence: 1 = Absent; 2 = Light; 3 = Heavy 1 1 Luster: 1 = Dull; 2 =Shiny 2 2 4. FLOWER: Type: 1 = Single; 2 = Semi-Double; 3 = Double 1 1Form: 1 = Flat; 2 = Cupped; 3 = Other 1 1 Shape: 1 = Round (PetalsOverlap); 2 = Intermediate; 3 = Star 1 1 (Petals Gapped) Flower Odor: 1= None; 2 = Mild; 3 = Strong 1 1 Pedicel Anthocyanin: 1 = Absent; 2 =Faint; 3 = Strong 1 1 Number Flowers per Plant 18 25 Flower Diameter(mm) 59 64 Orifice Size Including the Opening of the Corolla Tube (mm) 33 Petal Width At Widest Point (mm) 38 37 Petal Length From OutsideOrifice to Outer Edge (mm) 29 32 5. FLOWER COLORS: Petal Color ChartCode 2415C 2060C Orifice Color Chart Code 2425C 2063C 6. SEEDS (Mature(Dry) Seeds): Seed Set: 1 = None; 2 = Poor; 3 = Fair; 4 = Good; 5 =Excellent 4 4 Seed Coat Color: 1 = White; 2 = Tan; 3 = Brown; 4 = Black;4 4 5 = Other Seed Weight (mg/1000 Seeds) 1,641 1,944 *These are typicalvalues. Values may vary due to environment. Other values that aresubstantially equivalent are also within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of Line R2358Dand ‘PAS1305707’ CHARACTERISTIC R2358D ‘PAS1305707’ 1. OVERALL PLANTHABIT (at flowering stage): Species: 1 = C. roseus; 2 = Other 1 1Ploidy: 1 = Haploid; 2 = Diploid; 3 = Triploid; 4 = Tetrapioid 2 2 LifeCycle: 1 = Annual; 2 = Biennial; 3 = Perennial 1 1 Growth Habit: 1 =Determinate; 2 = Semi-determinate; 2 2 3 = Indeterminate Growth Form: 1= Upright; 2 = Semi-prostrate; 3 = Prostrate 1 1 Flowering: 1 = VeryEarly; 2 = Early; 3 = Mid-season; 4 = Late; 2 2 5 = Continuous Days fromPlanting to 50% Flowering 63 56 Length of Flowering Season in Days Untilfrost Until frost Plant Height at Maturity (cm) 18.8 23.7 Plant Width atMaturity (cm) 27.7 29.4 Plant Height Class: 1 = Extra Dwarf; 2 = Dwarf;3 = Semi-dwarf; 2 3 4 = Tall Plant Width Class: 1 = Compact; 2 =Semi-compact; 1 2 3 = Spreading/Lax 2. STEM: Profile: 1 = Straight; 2 =Zig-Zag 1 1 Branching Pattern: 1 = Single Stem; 2 = Few Branches; 3 =Many 3 3 Branches Stem Length from Base of Stem to Terminal Flower (cm)19.8 24.6 Number of Internodes Below First Branch 1 1 Number of FirstOrder Branches (From Main Stem) 12 14 Stem Anthocyanin: 1 = Absent; 2 =Along Veins Only; 3 = Solid 2 1 Coloration 3. FOLIAGE: Leaf Type: 1 =Simple; 2 = Compound 1 1 Leaf Margin: 1 = Entire; 2 = Serrate; 3 = Other1 1 Leaf Odor: 1 = None; 2 = Mild; 3 = Strong 1 1 Petiole Anthocyanin: 1= Absent; 2 = Mild; 3 = Strong 1 1 Leaf Shape: 1 = Lanceolate; 2 =Elliptic; 3 = Obovate; 4 = Ovate 2 2 Leaf Width (mm) 35.7 34.2 LeafLength (mm) 76.6 87.1 LEAF DORSAL SIDE: Leaf Color: 1 = Light Green; 2 =Medium Green; 3 = Dark Green; 3 3 4 = Other Color Chart Code  574C  574CPubescence: 1 = Absent; 2 = Light; 3 = Heavy 1 1 Luster: 1 = Dull; 2 =Shiny 1 1 LEAF VENTRAL SIDE: Leaf Color: 1 = Light Green; 2 = MediumGreen; 3 = Dark Green; 2 2 4 = Other _(——) _(——) Color Chart Code  371C 371C Pubescence: 1 = Absent; 2 = Light; 3 = Heavy 1 1 Luster: 1 = Dull;2 = Shiny 2 2 4. FLOWER: Type: 1 = Single; 2 = Semi-Double; 3 = Double 11 Form: 1 = Flat; 2 = Cupped; 3 = Other 1 1 Shape: 1 = Round (PetalsOverlap); 2 = Intermediate; 3 = Star 1 1 (Petals Gapped) Flower Odor: 1= None; 2 = Mild; 3 = Strong 1 1 Pedicel Anthocyanin: 1 = Absent; 2 =Faint; 3 = Strong 1 1 Number Flowers per Plant 15 18 Flower Diameter(mm) 60 59 Orifice Size Including the Opening of the Corolla Tube (mm) 33 Petal Width At Widest Point (mm) 38 38 Petal Length From OutsideOrifice to Outer Edge (mm) 29 29 5. FLOWER COLORS: Petal Color ChartCode 2063C 2415C Orifice Color Chart Code 2425C 2425C 6. SEEDS (Mature(Dry) Seeds): Seed Set: 1 = None; 2 = Poor; 3 = Fair; 4 = Good; 5 =Excellent 4 4 Seed Coat Color: 1 = White; 2 = Tan; 3 = Brown; 4 = Black;4 4 5 = Other Seed Weight (mg/1000 Seeds) 1,408 1,641 *These are typicalvalues. Values may vary due to environment. Other values that aresubstantially equivalent are also within the scope of the invention.

E. Distinguishing Characteristics of Hybrid ‘PAS1305707’ and ParentInbred Line R2358D

The closest commercial comparison for ‘PAS1305707’ of the presentinvention is believed to be ‘PAS1053045’, marketed as Valiant Lilac. Inaddition, ‘PAS1305707’ can be distinguished from its female parent,inbred line R2358D. Distinguishing characteristics were evaluated inboth a greenhouse trial grown in Guadalupe, Calif., and two field trialsgrown in Elburn, Ill.

For the Guadalupe, Calif. trial, plants were produced from seed andgrown in a glass-covered greenhouse under conditions comparable to thoseused in commercial practice. The plants were grown utilizing a soillessgrowth medium in one-gallon containers for 21 weeks. Greenhousetemperatures were maintained at approximately 75° F. to 85° F. (24° C.to 30° C.) during the day and approximately 65° F. to 68° F. (18° C. to20° C.) during the night. No supplemental lighting was provided.

In comparison, the plants from the Elburn, Ill. trials were producedfrom seed and grown in a glass-covered greenhouse under conditionscomparable to those used in commercial practice using trays having deep2⅜″×2⅜″ growing cells and a soilless growth medium. Plants weretransplanted to the field in early summer. Data was collected after 15weeks of outdoor growth for Trial 1, and after 12 weeks for Trial 2.

Table 3 illustrates that plants of ‘PAS1305707’ are significantlyshorter than plants of ‘PAS1053045’. In addition, Table 4 and Table 5show that ‘PAS1305707’ has significantly shorter and wider leaves thanplants of ‘PAS1053045’. Table 1 illustrates that flower petals of‘PAS1305707’ are a different color when compared to ‘PAS1053045’. Thistrait is consistent across all trials conducted.

As shown in Table 6, ‘PAS1305707’ has significantly longer leaves thaninbred line R2358D.

TABLE 3 Plant height differences between Hybrids ‘PAS1305707’ and‘PAS1053045’ ‘PAS1305707’ ‘PAS1053045’ Sample t Avg. Plant Avg. PlantSize Each Critical t Trial Height (cm) Height (cm) Variety α = .05Statistic P Value Elburn 1 41.0 +/− 3.1 45.9 +/− 2.2 10 2.1 4.1 6.8E−04Guadalupe 23.7 +/− 1.8 28.6 +/− 1.5 10 2.1 6.5 3.7E−06

TABLE 4 Leaf length differences between Hybrids ‘PAS1305707’ and‘PAS1053045’ ‘PAS1305707’ ‘PAS1053045’ Sample t Avg. Leaf Avg. Leaf SizeEach Critical t Trial Length (mm) Length (mm) Variety α = .05 StatisticP Value Elburn 1 61.6 +/− 5.8  77.6 +/− 7.7 25 2.0 8.3 6.9E−11 Guadalupe87.1 +/− 3.3 106.2 +/− 3.5 10 2.1 12.6 2.3E−10

TABLE 5 Leaf width differences between Hybrids ‘PAS1305707’ and‘PAS1053045’ ‘PAS1305707’ ‘PAS1053045’ Sample t Avg. Leaf Avg. Leaf SizeEach Critical t Trial Width (mm) Width (mm) Variety α = .05 Statistic PValue Elburn 1 28.5 +/− 2.3 24.4 +/− 3.0 25 2.0 −5.4 2.0E−06 Guadalupe34.2 +/− 1.4 29.1 +/− 1.5 10 2.1 −7.8 3.5E−07

TABLE 6 Leaf length differences between Hybrid ‘PAS1305707’ and R2358D‘PAS1305707’ R2358D Sample t Avg. Leaf Avg. Leaf Size Each Critical tTrial Length (mm) Length (mm) Variety α = .05 Statistic P Value Elburn 272.6 +/− 4.7 61.0 +/− 4.5 10 2.1 5.7 2.3E−05 Guadalupe 87.1 +/− 3.3 76.6+/− 3.7 10 2.1 6.7 2.6E−06

F. Breeding Catharanthus Plants

One aspect of the current invention concerns methods for producing seedof Catharanthus hybrid ‘PAS1305707’ involving crossing Catharanthuslines R2358D and M6948D. Alternatively, in other embodiments of theinvention, hybrid ‘PAS1305707’ or line R2358D may be crossed with itselfor with any second plant. Such methods can be used for propagation ofhybrid ‘PAS1305707’ or the Catharanthus line R23581D, or can be used toproduce plants that are derived from hybrid ‘PAS1305707’ or Catharanthusline R2358D. Plants derived from Catharanthus hybrid ‘PAS1305707’ orCatharanthus line R2358D may be used, in certain embodiments, for thedevelopment of new Catharanthus varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid ‘PAS1305707’ followed by multiplegenerations of breeding according to such well-known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g., colchicine treatment). Alternatively, haploid embryos may begrown into haploid plants and treated to induce chromosome doubling. Ineither case, fertile homozygous plants are obtained. In accordance withthe invention, any of such techniques may be used in connection with aplant of the invention and progeny thereof to achieve a homozygous line.

Backcrossing can be used to improve a variety, and may be used, forexample, to introduce a desired allele or trait into the plant geneticbackground of any plant that is sexually compatible with a plant of thepresent invention. Backcrossing transfers a specific desired trait fromone inbred or non-inbred source to a variety that lacks that trait. Thiscan be accomplished, for example, by first crossing a variety of adesired genetic background (recurrent parent) to a donor inbred(non-recurrent parent), which carries the appropriate allele or loci forthe desired trait(s) in question. The progeny of this cross are thenmated back to the recurrent parent, followed by selection in theresultant progeny for the desired trait to be transferred from thenon-recurrent parent. The process is repeated, for example for five ormore backcross generations with selection for the desired trait, theprogeny have the characteristic being transferred, but are like thesuperior parent for most or almost all other loci. The last backcrossgeneration would be selfed to give pure breeding progeny for the traitbeing transferred.

G. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those Catharanthus plants which are developed by aplant breeding technique called backcrossing or by genetic engineering,wherein essentially all of the morphological and physiologicalcharacteristics of a variety are recovered or conserved in addition tothe single locus introduced into the variety via the backcrossing orgenetic engineering technique, respectively. By essentially all of themorphological and physiological characteristics, it is meant that thecharacteristics of a plant are recovered or conserved that are otherwisepresent when compared in the same environment, other than an occasionalvariant trait that might arise during backcrossing, introduction of atransgene, or application of a genetic engineering technique.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalCatharanthus plant which contributes the locus for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental Catharanthus plant to which the locus or loci from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a plant is obtainedwherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent. The progeny thus have the characteristic being transferred, butare like the superior parent for most or almost all other loci. The lastbackcross generation can be selfed to give true-breeding progeny whenthe trait being transferred is introgressed into a true-breedingvariety.

The selection of a suitable recurrent parent therefore is an importantstep for a successful backcrossing procedure. The goal of a backcrossprotocol is to alter or substitute a single trait or characteristic inthe original variety. To accomplish this, a single locus of therecurrent variety is modified or substituted with the desired locus fromthe nonrecurrent parent, while retaining essentially all of the rest ofdesired genetic, and therefore the desired physiological andmorphological constitution of the original variety. The choice of theparticular nonrecurrent parent will depend on the purpose of thebackcross, one of the major purposes is to add some commerciallydesirable trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered and the geneticdistance between the recurrent and nonrecurrent parents. Althoughbackcrossing methods are simplified when the characteristic beingtransferred is a dominant allele, a recessive allele or an additiveallele (between recessive and dominant) may also be transferred. In thisinstance it may be necessary to introduce a test of the progeny todetermine if the desired characteristic has been successfullytransferred.

In one embodiment, progeny Catharanthus plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of the recurrent parent as determinedat the 5% significance level when grown in the same environmentalconditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the genetic background of the plants.In selecting a second plant to cross with Catharanthus hybrid‘PAS1305707’ or Catharanthus line R2358D for the purpose of developingnovel Catharanthus lines, it will typically be preferred to choose thoseplants which either themselves exhibit one or more selected desirablecharacteristics or which exhibit the desired characteristic(s) when inhybrid combination. Examples of desirable traits may include, inspecific embodiments, high flower yield, flower quality, high seedgermination, seedling vigor, disease resistance, and adaptability forsoil and climate conditions such as drought or heat. Consumer-driventraits, such as flower color, shape, and texture are other examples oftraits that may be incorporated into new lines of Catharanthus plantsdeveloped by this invention.

In other embodiments, the invention provides methods of vegetativelypropagating a Catharanthus plant of the present invention. Such a methodmay comprise the steps of: (a) collecting tissue capable of beingpropagated from said plant; and (b) propagating a Catharanthus plantfrom said tissue.

In still further embodiments, a plant of the invention is propagated byseed, wherein a plant may be used as either a female or a male parentfor producing progeny seed and plants.

Also provided are methods of producing a Catharanthus plant of thepresent invention, said method comprising introgressing a desired allelefrom a plant comprising the allele into a plant of a different genotype.In certain embodiments, such an allele may be inherited from orintrogressed into Catharanthus hybrid ‘PAS1305707’ or a progeny of anygeneration thereof comprising the allele.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,resistance to bacterial, fungal, or viral disease, or herbicide orinsect resistance. These comprise genes generally inherited through thenucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of Catharanthus plants for breeding is not necessarilydependent on the phenotype of a plant and instead can be based ongenetic investigations. Thus, in one embodiment, the invention providesthe genetic complement of a Catharanthus plant as described herein.“Genetic complement” as used herein refers to the aggregate ofnucleotide sequences, the expression of which sequences defines thephenotype of, in the present case, a Catharanthus plant, or a cell ortissue of that plant. A genetic complement thus represents the geneticmakeup of a cell, tissue or plant, and a hybrid genetic complementrepresents the genetic makeup of a hybrid cell, tissue or plant. Thegenetic complement of variety ‘PAS1305707’ may be identified by any ofthe many well-known techniques in the art. For example, one can utilizea suitable genetic marker which is closely genetically linked to a traitof interest. One of these markers can be used to identify the presenceor absence of a trait in the offspring of a particular cross, and can beused in selection of progeny for continued breeding. This technique iscommonly referred to as marker assisted selection.

Any other type of genetic marker or other assay which is able toidentify the relative presence or absence of a trait of interest in aplant can also be useful for breeding purposes. Procedures for markerassisted selection are well known in the art. Such methods will be ofparticular utility in the case of recessive traits and variablephenotypes, or where conventional assays may be more expensive, timeconsuming or otherwise disadvantageous. In addition, marker assistedselection may be used to identify plants comprising desirable genotypesat the seed, seedling, or plant stage, to identify or assess the purityof a cultivar, to catalog the genetic diversity of a germplasmcollection, and to monitor specific alleles or haplotypes within anestablished cultivar.

Types of genetic markers which could be used in accordance with theinvention include, but are not necessarily limited to, Simple SequenceLength Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 18:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR),Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858,specifically incorporated herein by reference in its entirety), andSingle Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiology, 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into a line.This molecular breeding-facilitated movement of a trait or traits into aline or variety may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the line orvariety by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into a line via this methodology. When thisline containing the additional loci is further crossed with anotherparental line to produce hybrid offspring, it is possible to thenincorporate at least eight separate additional loci into the hybrid.These additional loci may confer, for example, such traits as diseaseresistance, drought or heat tolerance, or a flower quality trait. In oneembodiment, each locus may confer a separate trait. In anotherembodiment, loci may need to be homozygous and exist in each parent lineto confer a trait in the hybrid. In yet another embodiment, multipleloci may be combined to confer a single robust phenotype of a desiredtrait.

In particular embodiments of the invention, marker assisted selection isused to increase the efficiency of a backcrossing breeding scheme forproducing a Catharanthus line comprising a desired trait. This techniqueis commonly referred to as marker assisted backcrossing (MABC). Thistechnique is well-known in the art and may involve, for example, the useof three or more levels of selection, including foreground selection toidentity the presence of a desired locus, which may complement orreplace phenotype screening protocols; recombinant selection to minimizelinkage drag; and background selection to maximize recurrent parentgenome recovery.

H. Plants Derived by Genetic Engineering

Various genetic engineering technologies have been developed and may beused by those of skill in the art to introduce traits in plants. Incertain aspects of the claimed invention, traits are introduced intoCatharanthus plants via altering or introducing a single genetic locusor transgene into the genome of a recited variety or progenitor thereof.Methods of genetic engineering to modify, delete, or insert genes andpolynucleotides into the genomic DNA of plants are well-known in theart.

In specific embodiments of the invention, improved Catharanthus linescan be created through the site-specific modification of a plant genome.Methods of genetic engineering include, for example, utilizingsequence-specific nucleases such as zinc-finger nucleases (see, forexample, U.S. Pat. Appl. Pub. No. 2011-0203012); engineered or nativemeganucleases; TALE-endonucleases (see, for example, U.S. Pat. Nos.8,586,363 and 9,181,535); and RNA-guided endonucleases, such as those ofthe CRISPR/Cas systems (see, for example, U.S. Pat. Nos. 8,697,359 and8,771,945 and U.S. Pat. Appl. Pub. No. 2014-0068797). One embodiment ofthe invention thus relates to utilizing a nuclease or any associatedprotein to carry out genome modification. This nuclease could beprovided heterologously within donor template DNA for templated-genomicediting or in a separate molecule or vector. A recombinant DNA constructmay also comprise a sequence encoding one or more guide RNAs to directthe nuclease to the site within the plant genome to be modified. Furthermethods for altering or introducing a single genetic locus include, forexample, utilizing single-stranded oligonucleotides to introduce basepair modifications in a Catharanthus plant genome (see, for exampleSauer et al., Plant Physiol, 170(4):1917-1928, 2016).

Methods for site-directed alteration or introduction of a single geneticlocus are well-known in the art and include those that utilizesequence-specific nucleases, such as the aforementioned, or complexes ofproteins and guide-RNA that cut genomic DNA to produce a double-strandbreak (DSB) or nick at a genetic locus. As is well-understood in theart, during the process of repairing the DSB or nick introduced by thenuclease enzyme, a donor template, transgene, or expression cassettepolynucleotide may become integrated into the genome at the site of theDSB or nick. The presence of homology arms in the DNA to be integratedmay promote the adoption and targeting of the insertion sequence intothe plant genome during the repair process through homologousrecombination or non-homologous end joining (NHEJ).

In another embodiment, genetic transformation may be used to insert aselected transgene into a plant of the invention or may, alternatively,be used for the preparation of transgenes which can be introduced bybackcrossing. Methods for the transformation of plants that are wellknown to those of skill in the art and applicable to many crop speciesinclude, but are not limited to, electroporation, microprojectilebombardment, Agrobacterium-mediated transformation and direct DNA uptakeby protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Nat. Biotechnol., 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Nat. Biotechnol., 3:629-635, 1985; U.S.Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance, or anyother gene of agronomic interest. Examples of constitutive promotersuseful for plant gene expression include, but are not limited to, thecauliflower mosaic virus (CaMV) P-35S promoter, which confersconstitutive, high-level expression in most plant tissues (see, e.g.,Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g.,Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol.Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV35S promoter, the enhanced 35S promoter (P-e35S);1 the nopaline synthasepromoter (An et al., Plant Physiol., 88:547, 1988); the octopinesynthase promoter (Fromm et al., Plant Cell, 1:977, 1989); and thefigwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No.5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where thepromoter sequence of P-FMV is duplicated in tandem; the cauliflowermosaic virus 19S promoter; a sugarcane bacilliform virus promoter; acommelina yellow mottle virus promoter; and other plant DNA viruspromoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., PlantPhysiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter,Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-bindingprotein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones,such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4)wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a Catharanthus plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a Catharanthus plant includeone or more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as environmental or stresstolerances, or any desirable changes in plant physiology, growth,development, morphology or plant product(s). For example, structuralgenes would include any gene that confers insect tolerance including butnot limited to a Bacillus insect control protein gene as described in WO99/31248, herein incorporated by reference in its entirety, U.S. Pat.No. 5,689,052, herein incorporated by reference in its entirety, U.S.Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference intheir entirety. In another embodiment, the structural gene can confertolerance to the herbicide glyphosate as conferred by genes including,but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPSgene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, hereinincorporated by reference in its entirety, or glyphosate oxidoreductasegene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporatedby reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, BirdBiotech. Gen. Engin. Rev., 9:207, et al., 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

I. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a genetic locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F1), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions or transgenes from one geneticbackground into another.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Plant Part: As used herein, a plant part refers to a part of a plant ofthe present invention. A plant part may be defined as comprising a cellof such plant, such as a cutting, a leaf, an ovule, pollen, a cell, aseed, a flower, an embryo, a meristem, a cotyledon, an anther, a root, aroot tip, a pistil, a stem, and a protoplast or callus derivedtherefrom.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing or genetic engineering ofa locus, wherein essentially all of the morphological and physiologicalcharacteristics of a Catharanthus variety are recovered in addition tothe characteristics of the single locus transferred into the variety viathe backcrossing technique or genetic engineering technique.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a Catharanthus plant by transformation.

Catharanthus plant: As used herein, Catharanthus refers to any plantfrom the genus Catharanthus, which may include but is not limited toCatharanthus pusillus, Catharanthus roseus, Catharanthus longifolius,Catharanthus ovalis, Catharanthus trichophyllus, and the like.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

J. Deposit Information

A deposit of Catharanthus hybrid ‘PAS1305707’ and parent line R2358D,disclosed above and recited in the claims, has been made with theAmerican Type Culture Collection (ATCC), 10801 University Blvd.,Manassas, Va. 20110-2209. The dates of the deposits made were ______,and ______, respectively. The accession numbers for those depositedseeds of Catharanthus hybrid ‘PAS1305707’ and inbred parent line R2358Dare ATCC Accession Number PTA-______, and ATCC Accession NumberPTA-______, respectively. Upon issuance of a patent, all restrictionsupon the deposits will be removed, and the deposits are intended to meetall of the requirements of 37 C.F.R. § 1.801-1.809. The deposits will bemaintained in the depository for a period of 30 years, or 5 years afterthe last request, or for the effective life of the patent, whichever islonger, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed is:
 1. A Catharanthus plant comprising at least a firstset of the chromosomes of Catharanthus line R2358D, a sample of seed ofsaid line having been deposited under ATCC Accession Number PTA-______.2. A Catharanthus seed that produces the plant of claim
 1. 3. The plantof claim 1, wherein the plant is a plant of said Catharanthus lineR2358D.
 4. The plant of claim 1, wherein said plant is a hybridCatharanthus plant.
 5. The seed of claim 2, wherein the seed is a seedof said Catharanthus line R2358D.
 6. The seed of claim 2, wherein theseed is a seed of a hybrid Catharanthus plant.
 7. The plant of claim 4,wherein the hybrid plant is a plant of Catharanthus hybrid ‘PAS1305707’,a sample of seed of said hybrid ‘PAS1305707’ having been deposited underATCC Accession Number PTA-______.
 8. The seed of claim 6, wherein theseed is a seed of Catharanthus hybrid ‘PAS1305707’, a sample of seed ofsaid hybrid ‘PAS1305707’ having been deposited under ATCC AccessionNumber PTA-______.
 9. A plant part of the plant of claim
 1. 10. Theplant part of claim 9, further defined as a flower, pollen, a leaf, anovule, an embryo, a seed, or a cell.
 11. A Catharanthus plant having allof the physiological and morphological characteristics of the plant ofclaim
 7. 12. A tissue culture of regenerable cells of the plant ofclaim
 1. 13. A Catharanthus plant regenerated from the tissue culture ofclaim
 12. 14. A method of vegetatively propagating a Catharanthus plant,the method comprising the steps of: (a) collecting tissue capable ofbeing propagated from the plant of claim 1; and (b) propagating aCatharanthus plant from said tissue.
 15. A method of introducing a traitinto a Catharanthus line, the method comprising: (a) utilizing as arecurrent parent the plant of claim 1 by crossing said plant with adonor plant that comprises a trait to produce F₁ progeny; (b) selectingan F₁ progeny that comprises the trait; (c) backcrossing the selected F₁progeny with the same line used as the recurrent parent in step (a) toproduce backcross progeny; (d) selecting a backcross progeny thatcomprises the trait and otherwise comprises the physiological andmorphological characteristics of the recurrent parent line used in step(a); and (e) repeating steps (c) and (d) three or more times to producea selected fourth or higher backcross progeny.
 16. A Catharanthus plantproduced by the method of claim
 15. 17. A method of producing a plantcomprising an added trait, the method comprising introducing a transgeneconferring the trait into the plant of claim
 1. 18. A Catharanthus plantproduced by the method of claim
 17. 19. A Catharanthus plant comprisingat least a first set of the chromosomes of Catharanthus line R2358D, asample of seed of said line having been deposited under ATCC AccessionNumber PTA-______, further comprising a transgene.
 20. The plant ofclaim 19, wherein the transgene confers a trait selected from the groupconsisting of male sterility, herbicide tolerance, insect resistance,pest resistance, disease resistance, and environmental stress tolerance.21. A Catharanthus plant comprising at least a first set of thechromosomes of Catharanthus line R2358D, a sample of seed of said linehaving been deposited under ATCC Accession Number PTA-______, furthercomprising a single locus conversion.
 22. The plant of claim 21, whereinthe single locus conversion confers a trait selected from the groupconsisting of male sterility, herbicide tolerance, insect resistance,pest resistance, disease resistance, and environmental stress tolerance.23. A method for producing a seed of a plant derived from at least oneof hybrid ‘PAS1305707’ or line R2358D, the method comprising the stepsof: (a) crossing the plant of claim 1 with itself or a secondCatharanthus plant; and (b) allowing seed of a hybrid ‘PAS1305707’- orline R2358D-derived Catharanthus plant to form.
 24. A method forproducing a seed of a hybrid ‘PAS1305707’- or line R2358D-derivedCatharanthus plant, the method comprising the steps of (a) producing ahybrid ‘PAS1305707’- or line R2358D-derived Catharanthus plant from aseed produced by crossing the plant of claim 1 with itself or adifferent Catharanthus plant; and (b) crossing the hybrid ‘PAS1305707’-or line R2358D-derived Catharanthus plant with itself or a differentCatharanthus plant to obtain seed of a further hybrid ‘PAS1305707’- orline R2358D-derived Catharanthus plant.
 25. The method of claim 24, themethod further comprising repeating said producing and crossing steps of(a) and (b) using the seed from said step (b) for producing the plantaccording to step (a) for at least one generation to produce a seed ofan additional hybrid ‘PAS1305707’- or line R2358D-derived Catharanthusplant.
 26. A plant part of the plant of claim
 7. 27. The plant part ofclaim 26, further defined as a flower, pollen, a leaf, an ovule, anembryo, a seed, or a cell.
 28. A method of producing a Catharanthusseed, the method comprising: (a) obtaining the plant of claim 1, whereinthe plant has been cultivated to maturity; and (b) collecting aCatharanthus seed from the plant.